α-细辛醚对无镁细胞外液培养海马神经元构建难治性癫痫细胞模型的影响

Effect of α-asarone on construction of intractable epilepsy cell models by culturing hippocampal neurons with magnesium-free extracellular solution

背景:无镁细胞外液处理培养的海马神经元可诱导产生反复自发性癫痫样放电,该模型可作为临床难治性癫痫细胞模型。目的:探讨难治性癫痫细胞模型中α-细辛醚对神经元的保护作用。方法:分离培养新生24h内的SD大鼠海马神经元,取经鉴定的海马神经元,加入含终浓度为7.5,15,30,60,120mg/Lα-细辛醚的维持培养液培养4h后,换为无镁液建立难治性癫痫细胞模型,3h后恢复含α-细辛醚的维持培养基继续培养24h,MTT法检测海马神经元活力。结果与结论:经无镁细胞外液培养后,海马神经元的活力显著降低(P<0.01),α-细辛醚处理后,海马神经元的活力显著升高(P<0.01),且随α-细辛醚浓度的增加,海马神经元的相对活力升高。说明α-细辛醚可以抑制难治性癫痫细胞模型中的神经元损伤,发挥神经元保护作用,且具有剂量依赖性。

BACKGROUND：The hippocampal neurons exposed to magnesium-free extracellular solution could develop and express spontaneous recurrent epileptionform discharges.The model can be used as intractable epilepsy cell models in the clinic.OBJECTIVE：To investigate the protective effect of α-asarone on intractable spilepsy cell models.METHODS：The rat hippocampal neurons were harvested within 24 hours after rat birth.The neurons were identified by immunochemistry and cultured with 7.5,15,30,60,120 mg/L α-asarone.After 4 hours,the hippocampal neurons were exposed to magnesium-free extracellular solution to establish intractable spilepsy cell models.After 3 hours,the neurons were cultured with medium containing α-asarone for 24 hours.The vitality of hippocampal neurons was detected by thiazolyl blue tetrazolium bromide.RESULTS AND CONCLUSION：Following culture with magnesium-free extracellular solution,the vitality of hippocampal neurons was significantly decreased（P 0.01）,and after culture with α-asarone,the vitality of hippocampal neurons was relatively increased.These results demonstrated that α-asarone can alleviate the damage to neurons in intractable spilepsy,exhibiting a protective effect on neurons in a dose-dependent manner.